The main purpose of this paper is to review the various methods used for evaluation of fluoride retention in saliva, plaque, and enamel following application of topical anti-caries treatments such as F dentifrices and F mouthwashes. Such methods monitor delivery of fluoride to the site of action, the mouth, and so can be regarded as assessing potential for treatment action. It is concluded that intra-oral fluoride measurements are appropriate to support bioequivalence claims for anti-caries treatments, provided that particular chosen methods have been calibrated against clinical data. Studies purporting to show superiority are of interest mechanistically, but links to caries are not sufficiently understood to define superiority claims. A wide variety of methods has been used for determination of the fluoride content of enamel. Of these, well-established methods such as the micro-drill and acid-etch procedures are appropriate for routine comparative testing, whereas sophisticated instrumental techniques such as SIMS are more appropriate for detailed mechanistic studies. Intra-oral pH measurements are also relevant to many topical treatments. Single-site determinations in plaque are preferred, but for comparative studies non-specific determinations may be adequate.